Malassezia globosa Activates NLRP3 Inflammasome in Human Sebocytes

Abstract

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NLRP3 regulates CIITA/MHC II axis and interferon‐γ‐inducible chemokines in Malassezia globosa‐infected keratinocytes

“…M. globosa ‐type strain CBS9597 was obtained from the Institute of Dermatology, Chinese Academy of Medical Sciences and Peking Union Medical College (Nanjing, China). The yeasts were cultivated in modified Leeming–Notman broth for 7 days at 32°C, harvested by centrifugation at 3000 × g for 3 min, washed thrice in PBS and diluted in medium 30 . Heat‐inactivated (HI)‐ M. globosa was prepared by incubation at 95°C for 15 min.…”

“…HaCaT cells were pretreated with cyD for 1 h, followed by coincubation with 30 MOI M. globosa or HI M. globosa for 24 h. The nuclear and cytoplasmic protein fractions were extracted using Nuclear and Cytoplasmic Protein Extraction Kit (Beyotime) according to the manufacturer's protocol. According to our previous experiment, 30 the blots were incubated with primary antibodies NLRP3, ASC, cleaved caspase‐1, cleaved IL‐1β, CBP, CIITA, STAT1, MHC II and GAPDH at a 1:1000 dilution and then with HRP‐conjugated secondary antibodies and visualised by enhanced chemiluminescence.…”

“…HaCaT cells were cultured in confocal Petri dishes and treated with 30 MOI M. globosa or other agents for 24 h. Immunofluorescence for NLRP3, ASC, caspase‐1 and CIITA was performed as previously described 30 …”

“…The yeasts were cultivated in modified Leeming-Notman broth for 7 days at 32°C, harvested by centrifugation at 3000 × g for 3 min, washed thrice in PBS and diluted in medium. 30 Heat-inactivated (HI)-M. globosa was prepared by incubation at 95°C for 15 min.…”

“…qRT-PCR was performed as described previously (Li et al, 2022). CCL1, CCL5, CCL22, CXCL8, CXCL10, CXCL11, CXCL1, CXCL9, CXCL1 and CCL2 primers (Table S2) were synthetised by Sangon.…”